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Leucine promotes leptin receptor expression in mouse C2C12 myotubes through the mTOR pathway

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Abstract

Leptin plays a critical role in regulating muscle protein metabolism by binding with leptin receptors in a 1:1 stoichiometry. However, the role for leucine in the regulation of leptin receptor expression in muscle has not been investigated. The present study was conducted to test the hypothesis that leucine regulates leptin receptor levels in C2C12 myotubes. Cells were cultured in the presence of DMEM/F12 medium containing supplemental 0 or 5 mM l-leucine. Leptin receptor expression by C2C12 myotubes peaked at 2 h post-supplementation. Additionally, leucine stimulated leptin receptor expression at both mRNA and protein levels in a dose-dependent manner. Furthermore, leucine enhanced the phosphorylation of mammalian target of rapamycin (mTOR). Addition of rapamycin (an inhibitor of mTOR) to culture medium completely suppressed leucine-induced activation of mTOR and inhibited leucine-stimulated leptin receptor production. These results indicate that leucine affects leptin receptor expression in muscle cells via the mTOR signaling pathway.

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References

  1. Zhang Y, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM (1994) Positional cloning of the mouse obese gene and its human homologue. Nature 372:425–432

    Article  PubMed  CAS  Google Scholar 

  2. Maffei M, Halaas J, Ravussin E, Pratley RE, Lee GH, Zhang Y, Fei H, Kim S, Lallone R, Ranganathan S, Kern PA, Freidman JM (1995) Leptin levels in human and rodent: measurement of plasma leptin and ob RNA in obese and weight reduced subjects. Nat Med 1:1155–1161

    Article  PubMed  CAS  Google Scholar 

  3. Schwartz MW, Baskin DG, Kaiyala KJ, Woods SC (1999) Model for the regulation of energy balance and adiposity by the central nervous system. Am J Clin Nutr 69:584–596

    PubMed  CAS  Google Scholar 

  4. Ramsay TG (2003) Porcine leptin inhibits protein breakdown and stimulates fatty acid oxidation in mouse C2C12 myotubes. J Anim Sci 81:3046–3051

    PubMed  CAS  Google Scholar 

  5. Carbó N, Ribas V, Busquets S, Alvarez B, Lpez-Soriano FJ, Argils JM (2000) Short-term effects of leptin on skeletal muscle protein metabolism in the rat. J Nutr Biochem 11:431–435

    Article  PubMed  Google Scholar 

  6. Lamoov D, Zeman M (2001) Effect of leptin and insulin on chick embryonic muscle cells and hepatocytes. Physiol Res 50:183–189

    Google Scholar 

  7. Tartaglia LA (1997) The leptin receptor. J Biol Chem 272:6093–6096

    PubMed  CAS  Google Scholar 

  8. Tartaglia LA, Dembski M, Weng X, Deng N, Culpepper J, Devos R, Richards GJ, Campfield LA, Clark FT, Deeds J, Muir C, Sanker S, Moriarty A, Moore KJ, Smutko JS, Mays GG, Wool EA, Monroe CA, Tepper RI (1995) Identification and expression cloning of a leptin receptor, OB-R. Cell 83:1263–1271

    Article  PubMed  CAS  Google Scholar 

  9. Chua SC, Chung WK, Wu-Peng XS, Zhang Y, Liu SM, Tartaglia L, Leibel RL (1996) Phenotypes of mouse diabetes and rat fatty due to mutations in the OB (leptin) receptor. Science 271:994–996

    Article  PubMed  CAS  Google Scholar 

  10. Hegyi K, Fülöp K, Kovcás K, Tóth S, Falus A (2004) Leptin-induced signal transduction pathways. Cell Biol Int 28:159–169

    Article  PubMed  CAS  Google Scholar 

  11. Huang L, Wang ZW, Li C (2001) Modulation of circulating leptin levels by its soluble receptor. J Biol Chem 276:6343–6349

    Article  PubMed  CAS  Google Scholar 

  12. Crozier SJ, Kimball SR, Emmert SW, Anthony JC, Jefferson LS (2005) Oral leucine administration stimulates protein synthesis in rat skeletal muscle. J Nutr 135:376–382

    PubMed  CAS  Google Scholar 

  13. Han B, Tong JF, Zhu MJ, Ma CW, Du M (2008) Insulin-like growth factor-1 (IGF-1) and leucine activate pig myogenic satellite cells through mammalian target of rapamycin (mTOR) pathway. Mol Reprod Dev 75:810–817

    Article  PubMed  CAS  Google Scholar 

  14. Du M, Shen QW, Zhu MJ, Ford SP (2007) Leucine stimulates mammalian target of rapamycin signaling in mouse C2C12 myoblasts in part through inhibition of adenosine monophosphate-activated protein kinase. J Anim Sci 85:919–927

    Article  PubMed  CAS  Google Scholar 

  15. Suryawan A, Jeyapalan AS, Orellana RA, Wilson FA, Nquyen HV, Davis TA (2008) Leucine stimulates protein synthesis in skeletal muscle of neonatal pigs by enhancing mTORC1 activation. Am J Physiol Endocrinol Metab 295:E868–E875

    Article  PubMed  CAS  Google Scholar 

  16. Lynch CJ, Gern B, Lloyd C, Hutson SM, Eicher R, Vary TC (2006) Leucine in food mediates some of the postprandial rise in plasma leptin concentrations. Am J Physiol Endocrinol Metab 291:E621–E630

    Article  PubMed  CAS  Google Scholar 

  17. Roh C, Han J, Tzatsos A, Kandror KV (2003) Nutrient-sensing mTOR-mediated pathway regulates leptin productin in isolated rat adipocytes. Am J Physiol Endocrinol Metab 284:E322–E330

    PubMed  CAS  Google Scholar 

  18. Devos R, Guisez Y, Van der Heyden J, White DW, Kalai M, Fountoulakis M, Plaetinck G (1997) Ligand-independent dimerization of the extracellular domain of the leptin receptor and determination of the stoichiometry of leptin binding. J Biol Chem 272:18304–18310

    Article  PubMed  CAS  Google Scholar 

  19. Kimball SR (2002) Regulation of global and specific mRNA translation by amino acids. J Nutr 132:883–886

    PubMed  CAS  Google Scholar 

  20. Fafournoux P, Bruhat A, Jousse C (2000) Amino acid regulation of gene expression. Biochem J 351:1–12

    Article  PubMed  CAS  Google Scholar 

  21. Lai CH, Yin JD, Li DF, Zhao LD, Qiao SY, Xing JJ (2005) Conjugated linoleic acid attenuates the production and gene expression of proinflammatory cytokines in weaned pigs challenged with lipopolysaccharide. J Nutr 135:239–244

    CAS  Google Scholar 

  22. Zhou X, Li DF, Yin JD, Ni JJ, Dong B, Zhang JX, Du M (2007) CLA differently regulates adipogenesis in stromal vascular cells from porcine subcutaneous adipose and skeletal muscle. J Lipid Res 48:1701–1709

    Article  PubMed  CAS  Google Scholar 

  23. Kimball SR, Jefferson LS (2006) Signaling pathways and molecular mechanisms through which branched-chain amino acids mediate translational control of protein synthesis. J Nutr 136:227S–231S

    PubMed  CAS  Google Scholar 

  24. Drenan RM, Liu X, Bertram PG, Zheng XF (2004) FKBP12-rapamycin-associated protein or mammalian target of rapamycin (FRAP/mTOR) localization in the endoplasmic reticulum and the golgi apparatus. J Biol Chem 279:772–778

    Article  PubMed  CAS  Google Scholar 

  25. Koros C, Boukouvalas G, Gerozissis K, Kitraki E (2009) Fat diet affects leptin receptor levels in the rat cerebellum. Nutrition 25:85–87

    Article  PubMed  CAS  Google Scholar 

  26. Alonso A, Fernández R, Moreno M, Ordóñez P, Díaz F, González C (2007) Leptin and its receptor are controlled by 17β-estradiol in peripheral tissue of ovariectomized rats. Exp Biol Med 232:542–549

    CAS  Google Scholar 

  27. Chen XJ, Li DF, Yin JD, Ding YH, Zhang HL, Zhang HW, Yi GF (2006) Regulation of dietary energy level and oil source on leptin and its long form receptor mRNA expression of the adipose tissue in growing pigs. Domest Anim Endocrinol 31:269–283

    Article  PubMed  CAS  Google Scholar 

  28. Berti L, Gammeltoft S (1999) Leptin stimulates glucose uptake in mouse C2C12 muscle cells by activation of ERK2. Mol Cell Endocrinol 157:121–130

    Article  PubMed  CAS  Google Scholar 

  29. Tajmir P, Kwan JJ, Kessas M, Mozammel S, Sweeney G (2003) Acute and chronic leptin treatment mediate contrasting effects on signaling, glucose uptake, and GLUT4 translocation in L6-GLUT4myc myotubes. J Cell Physiol 197:122–130

    Article  PubMed  CAS  Google Scholar 

  30. Tomiya T, Nishikawa T, Inoue Y, Ohtomo N, Ikeda H, Tejima K, Wtanabe N, Tanoue Y, Omata M, Fujiwara K (2007) Leucine stimulates HGF production by hepatic stellate cells through mTOR pathway. Biochem Biophy Res Commun 358:176–180

    Article  CAS  Google Scholar 

  31. Deldicque L, Sanchez Canedo C, Horman S, De Potter I, Bertrand L, Hue L, Francaux M (2008) Antagonistic effects of leucine and glutamine on the mTOR pathway in myogenic mouse C2C12 cells. Amino Acids 35:147–155

    Article  PubMed  CAS  Google Scholar 

  32. Ijichi C, Matsumura T, Tsuji T, Eto Y (2003) Branched-chain amino acids promote albumin synthesis in rat primary hepatocytes through the mTOR signal transduction system. Biochem Biophy Res Commun 303:59–64

    Article  CAS  Google Scholar 

  33. Lynch CJ (2001) Role of leucine in the regulation of mTOR by amino acids: revelations from structure-activity studies. J Nutr 131:861S–865S

    PubMed  CAS  Google Scholar 

  34. Anthony JC, Yoshizawa F, Anthony TG, Vary TC, Jefferson LS, Kimball SR (2000) Leucine stimulates translation initiation in skeletal muscle of postabsorptive rat via a rapamycin-sensitive pathway. J Nutr 130:2413–2419

    PubMed  CAS  Google Scholar 

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Acknowledgments

This study was financially supported by grants from the National Science Foundation of China (30525029). Special thanks to Professor Malcolm Watford from the Rutgers University and Professor Guoyao Wu from the Texas A&M University for editing the manuscript.

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Correspondence to Shiyan Qiao.

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Mao, X., Zeng, X., Wang, J. et al. Leucine promotes leptin receptor expression in mouse C2C12 myotubes through the mTOR pathway. Mol Biol Rep 38, 3201–3206 (2011). https://doi.org/10.1007/s11033-010-9992-6

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  • DOI: https://doi.org/10.1007/s11033-010-9992-6

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